HSV-1/2; obesogen; stool softener; sulfated surfactant

Inactivation of HSV [1]
The in vitro inactivations of HSV by Docusate were compared with another surfactant, SDS. Various concentrations of testing drug were pre-mixed with HSV-1 or HSV-2 and incubated at 37 °C for 1 h. Antiviral effects were then determined by plaque reduction assays (Fig. 1). Inactivation of HSV-1 by Docusate along with SDS is shown in Fig. 1A and that of HSV-2 in Fig. 1B. Results showed that at a concentration of 0.005%, docusate completely inactivated both HSV-1 and HSV-2 after pre-mixing for 1 h (EC90–100=0.005%). In contrast, although this concentration was also effective for SDS, it caused 50% cell death (toxic to cells). At concentration of 0.01%, both docusate and SDS were shown to be toxic to the cells causing 50–100% cell death (data not shown). In these experiments, docusate was slightly more effective against HSV-1 than HSV-2, and slightly more potent and less toxic than SDS at other concentrations (Fig. 1).

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The kinetics of inactivation of HSV by Docusate [1]
To examine the kinetics of inactivation of HSV, docusate at various concentrations was mixed with HSV-2 and incubated at 37 °C in a time-course, with endpoints determined by plaque reduction assay. Results (Fig. 2) showed that docusate had a slight inactivation effect upon HSV-2 at concentrations of 0.001 and 0.0025% after 4 h incubation. However, inactivation reached 50% at time 0, almost 90% (EC90) after 30 min, and 100% after 1 h incubation at a concentration of 0.005%. A concentration of 0.01% of docusate was found to be toxic to cells.

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Short and long incubation cytotoxicities [1]
Microbicidal effectiveness of surface active agents such as Docusate against enveloped viruses suggests a potentially disruptive effect on cellular membranes. Accordingly, experiments were carried out to evaluate relative cytotoxicities of docusate over time. Cytotoxicities of docusate were compared with SDS and measured using Vero cells. Trypan blue exclusion (data not shown) and uptake of neutral red dye were then used to determine the viabilities of cells after incubations with different concentrations of docusate, and SDS. Results were consistent between assays (data not shown). For short term incubation, cells were exposed to docusate for 1 (Fig. 3A) and 6 h (data not shown), whereas for long term incubation, cells were exposed to docusate for 2 (data not shown) and 3 days (Fig. 3B). As can be seen from these figures, after 1 h exposure to docusate and SDS, minimal cytotoxicity of docusate to Vero cells was observed even at concentration of 0.01%. Cytotoxicity was increased after 6 h exposure (data not shown). Concentration at 0.01% was again found to be toxic to Vero cells after 3 days incubation. Thus, the cytotoxicity of docusate is time- and dose-dependent. Furthermore, docusate was slightly less cytotoxic than SDS. CC50 (cytotoxic concentration giving 50% of cell death) of docusate after 2 days incubation was approximately 0.01% and that of SDS was approximately 0.005%.

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Antiviral activity of Docusate determined by pre-treated cells [1]
Some compounds can be internalized into cells or bound to the cellular membranes to exert antiviral effects. This experiment was designed to examine the effect of Docusate on pre-treated cells. Two approaches were employed. First, we pre-incubated Vero cells with docusate and then infected these cells with HSV. In the second, we pre-incubated cells with docusate and then washed it off three times with PBS. The results, together with comparison of SDS, were summarized in Table 1. Cells pre-treated with docusate at a concentration of 0.005%, showed virus infection to be reduced by 45%, and 35% after docusate was removed by three washes. However, this concentration was not able to completely stop virus infection.

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Effect of Docusate on HSV-infected cells [1]
To examine the effect of Docusateon HSV-infected cells, cells were first infected with HSV-2 and then treated with different concentrations of docusate following adsorption by inclusion in the methylcellulose overlay. The results (Table 2) indicated that docusate only had slight inhibitory effect on HSV-infected cells, by approximately 30% plaque reduction at 0.005% of concentration.

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Effect of Docusate on HSV-drug-resistant mutants [1]
To test the antiviral effect of Docusate on the drug-resistant HSV strain, delta 333 virus was pre-mixed with different doses of docusate at 37 °C for 1 h. The antiviral effect was subsequently determined by plaque reduction assay. The results showed that docusate at concentration of 0.005% could completely inactivate delta 333 after incubation at 37 °C for 1 h (Table 3). Although SDS was slightly less efficacious in this assay, it was also capable of inactivating the resistant strain, delta 333. [1]
The effect of Docusate on the polymerase mutant virus was examined by pre-mixing docusate at various doses with 615.8 virus for 1 h at 37 °C. The antiviral activity against polymerase mutant virus was then determined by plaque reduction assay. The results showed that docusate at concentration of 0.005% could inactivate approximately 98% of polymerase mutant 615.8 after incubation at 37 °C for 1 h (Table 4), whereas SDS was less efficacious at this concentration.

Evidence indicates that obesity can be promoted by chemical 'obesogens' that drive adiposity, hunger, inflammation and suppress metabolism. Docusate/Dioctyl sodium sulfosuccinate (DOSS), a lipid emulsifier and candidate obesogen in vitro, is widely used in processed foods, cosmetics and as stool softener medicines commonly used during pregnancy. In vivo testing of Docusate/DOSS was performed in a developmental origins of adult obesity model. Pregnant mice were orally administered vehicle control or Docusate/DOSS at times and doses comparable to stool softener use during human pregnancy. All weaned offspring consumed only standard diet. Adult male but not female offspring of DOSS-treated dams showed significantly increased body mass, overall and visceral fat masses, and decreased bone area. They exhibited significant decreases in plasma adiponectin and increases in leptin, glucose intolerance and hyperinsulinemia. Inflammatory IL-6 was elevated, as was adipose Cox2 and Nox4 gene expressions, which may be associated with promoter DNA methylation changes. Multiple significant phospholipid/sterol lipid increases paralleled profiles from long-term high-fat diet induced obesity in males. Collectively, developmental DOSS exposure leads to increased adult adiposity, inflammation, metabolic disorder and dyslipidemia in offspring fed a standard diet, suggesting that pharmaceutical and other sources of DOSS taken during human pregnancy might contribute to long-term obesity-related health concerns in offspring [2].

Virus plaque reduction assay [1]
Antiviral effects of Docusate were determined by modified plaque reduction assays. Confluent cells were washed with PBS and subsequently infected with HSV for 1 h at 37 °C. After viral inoculum was removed, the infected cells were washed with PBS and overlaid with 0.5% methylcellulose in culture medium. Cells were incubated at 37 °C for 2 days for HSV-2 infection and 3 days for HSV-1. When plaque size was adequate, cells were fixed with 10% formalin and subsequently stained with 0.5% crystal violet. [1]
All data were generated from duplicate or triplicate wells in two or three independent experiments. Mean plaque counts are shown in the tables and figures. Effects of compounds at varying concentrations were expressed as percentage of control (the mean plaque counts in drug treated wells/the mean plaque counts in control wells).

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Effect of Docusate on HSV drug-resistant strains [1]
The antiviral effect of Docusate on HSV-2 thymidine kinase mutant Delta 333 and HSV-1 DNA polymerase mutant 615.8 was performed in a similar way as above (see Section 2.4)

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Inactivation of HSV by Docusate [1]
HSV G and F strains were diluted to 200 pfu/ml with 5% MEM, respectively. Docusate and SDS were diluted to 2× final concentrations (final concentrations: 0, 0.0005, 0.001, 0.0025, 0.005 and 0.01%, respectively) with 5% MEM. Equal volumes of diluted virus and drugs were mixed and incubated at 37 °C water-bath for 1 h. One milliliter of the mixture was then used to infect confluent Vero cells in 6-well plates at 37 °C for 1 h. After infection, viral inoculum was removed and the cells washed with PBS. The cells were subsequently overlaid with 1.5 ml of 0.5% methylcellulose in culture medium for plaque assay. [1]
To examine the kinetics of inactivation of HSV by Docusate, 200 pfu/ml of HSV-2 was pre-mixed with 2× final concentrations of docusate (final concentrations: 0, 0.001, 0.0025, 0.005 and 0.01%, respectively) in 5% MEM and incubated at 37 °C for 0, 15, 30, 60, 120, and 240 min. At each time point, the treated mixture was used to infect confluent Vero cells. After viral inoculum was removed, the cells were covered with methylcellulose for plaque assay.

Docusate Stock solutions (1%, w/v) was prepared by dissolving them in sterile deionized distilled water and stored at room temperature.

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Cytotoxicity of Docusate [1]
The cytotoxicity of Docusate was examined using Vero cells with the neutral red uptake assay described by Schmidt and Korba (2000). Culture medium was removed from confluent Vero cells in 24-well plates. The cells were then washed once with PBS. One milliliter of culture medium containing Docusate or SDS at concentrations of 0, 0.001, 0.0025, 0.005, and 0.01% were added to each well. Cells were incubated at 37 °C for 1 h, 6 h, 2 and 3 days. At each time point, the medium was removed and cells washed with PBS. Five hundred microlitres of 0.01% neutral red (in PBS) was added to each well, and the samples were incubated at 37 °C for 30 min. The dye was then removed and the cells washed twice with 1 ml PBS per well. The dye was extracted by addition of 500 μl of 50% ethanol/1% glacial acetic acid in PBS to each well and incubated at room temperature for 15 min with gentle shaking at 120–150 rpm. Then 200 μl of extracted dye from each well was put into 96-well plate and the absorbance at 550 nm was read on an ELISA reader. [1]
Long-term cytotoxicity of docusate was also examined using trypan blue exclusion assay. Confluent Vero cells were washed with PBS and incubated with culture medium containing various concentrations of Docusate or SDS at 37 °C for 3 days. After culture medium was removed, the cells were washed once with PBS, trypsinized with Versene/1× trypsin and resuspended in 1 ml of 5% MEM culture medium. One hundred microlitres of cell suspension was mixed with 500 μl trypan blue solution. The viable cells were counted using a hemacytometer.

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Pre-treatment of cells [1]
Confluent Vero cells in 6-well plates were washed with PBS. One milliliter culture medium containing different concentrations of Docusate or SDS was added to each well and the cells were incubated at 37 °C for 1 h. Following pre-incubation of the cells with Docusate, two treatments were performed, one set of plates was directly infected with 100 pfu per well of HSV-2; another set of plates was washed three times with PBS, and then infected with 100 pfu per well of HSV-2. Both infections were incubated at 37 °C for 1 h with tilting every 10 min. After viral inoculum was removed, the infected cells were covered with methylcellullose for plaque assay.

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Effect of Docusate on HSV-infected cells [1]
Confluent Vero cells were washed with PBS and then infected with 100 pfu per well of HSV-2 at 37 °C for 1 h. Following removal of viral inoculum, infected cells were washed once with PBS and covered with 0.5% methylcellulose containing either Docusate or SDS at various concentrations for plaque assay.

Pregnant dams (n = 3–6 per group) from E11.5 through weaning were provided a with water bottles containing either vehicle control (0.5% carboxymethylcellulose; CMC) or CMC plus 31.25 μg/mL Docusate/DOSS, both prepared in autoclaved 18MOhm water to simulate DOSS use during human pregnancy. Stool softeners (e.g. Docusate, Colace, etc.), which are essentially made of Docusate/DOSS, are accepted as safe prescription drugs and used by the medical community as the standard of care to treat constipation during pregnancy and breastfeeding. CMC was used to increase palatability of the DOSS solution and has been used as a vehicle control in other obesogen dosing studies. CMC is also commonly found as a component in DOSS-containing stool softeners. The DOSS dosage (31.25 μg/mL) correlates to the dose received by a pregnant woman (88.5 kg average weight, with a daily Docusate sodium stool softener dosage at 5.6 mg/kg = 500 mg DOSS) assuming that an adult mouse weighing 25 g drinks about 4 mL per day. The DOSS dosage given to pregnant dams (31.25 µg/mL) was more than 70-fold less than the no-observed-adverse-effect-level (NOAEL) for maternal DOSS toxicity and teratogenicity set for mice and rats (400 mg/kg) previously. The timing of exposure was based on the incidence of constipation in women becoming elevated from mid-gestation onward. Stock bottles of each solution were prepared as needed (usually weekly). A 1% CMC solution in autoclaved water and a 62.5 μg/mL solution of DOSS in autoclaved water were prepared and then mixed together in a ratio of 1:1 to achieve the final desired concentration for treatment. Autoclaved water was used to dilute the 1% CMC solution 1:1 to achieve the desired concentration of 0.5% CMC for the vehicle control. Only litters with 5–8 pups were used for analysis to control for nutrient availability in utero and potential effects of DOSS on litter size. Previous studies indicated that there was no adverse effect on litter size using more than 70-fold higher doses of DOSS. Dosing experiments were repeated three times and control vs. treatment animals from different experiments were grouped for analyses. Investigators were blinded as to the control and treated group samples.

Effects During Pregnancy and Lactation
◈ What is Sodium docusate?
Sodium docusate is a stool softener used to treat constipation (fewer than three bowel movements per week or difficulty defecating). Many products contain Sodium docusate. Some brand names include Colace®, Correctol®, Docusate®, Surfak®, and Soflax®. Sometimes, when people find out they are pregnant, they consider changing how they take the medication or even stopping it entirely. However, it is essential to consult your healthcare provider before changing your medication. Your healthcare provider can discuss with you the benefits of treating your condition and the risks of not treating it during pregnancy.
◈ I take Sodium docusate. Will taking Sodium docusate affect my pregnancy?
It is currently unclear whether Sodium docusate affects pregnancy.
◈ Does taking Sodium docusate increase the risk of miscarriage?
Miscarriage is common and can occur in any pregnancy for a variety of reasons. There is currently no research indicating that Sodium docusate increases the risk of miscarriage. Generally, only a very small amount of Sodium docusate is expected to enter the bloodstream and reach the developing fetus.
◈ Does taking Sodium docusate increase the risk of birth defects?
There is a 3-5% risk of birth defects in each pregnancy. This is called background risk. Current information suggests that, when used as directed, Sodium docusate is unlikely to increase the incidence of birth defects above the background risk.
◈ Does taking Sodium docusate during pregnancy increase the risk of other pregnancy-related problems?
When used at the recommended dose, Sodium docusate is unlikely to cause problems during pregnancy, such as preterm birth (delivery before 37 weeks of gestation) or low birth weight (birth weight less than 5 pounds 8 ounces [2500 grams]). Using Sodium docusate above the recommended dose can lower the magnesium level in the blood. There has been one reported case of neonatal hypomagnesemia associated with excessive Sodium docusate use by the mother. The infant's main symptom was irritability, which disappeared on the second day after birth. There are currently no reports of problems related to taking Sodium docusate at the recommended dose during pregnancy.
◈ Will taking Sodium docusate during pregnancy affect a child's future behavior or learning abilities?
Currently, there are no studies exploring whether Sodium docusate causes behavioral or learning problems in children.
◈ Breastfeeding while taking Sodium docusate:
Currently, there are no studies on taking Sodium docusate while breastfeeding. Sodium docusate is not easily absorbed by the gastrointestinal tract, so it is unlikely to be present in large quantities in breast milk. There has been one report of an infant experiencing diarrhea due to the presence of both Sodium docusate and another drug called dananthrone in breast milk. There are currently no other reports of problems in breastfed children due to the presence of Sodium docusate in breast milk. If you suspect your infant has any symptoms (such as diarrhea), contact your child's healthcare provider. Be sure to consult your healthcare provider about all questions regarding breastfeeding.
◈ If men take Sodium docusate, will it affect fertility (the ability to impregnate a partner) or increase the risk of birth defects?
Currently, there are no studies exploring whether Sodium docusate affects male fertility or increases the risk of birth defects (above background risk). Generally, medications that the father or sperm donor is exposed to are unlikely to increase the risk of pregnancy. For more information, please refer to MotherToBaby's "Paternal Exposure" information sheet at https://mothertobaby.org/fact-sheets/paternal-exposures-pregnancy/.

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23673837trattLD50toralt1900 mg/kgt Journal of the American Pharmaceutical Association, 13(469), 1962
23673837trattLD50tintraperitonealt590 mg/kgt Vascular: STRICTIRA; CJAMGES OM VESSE; St. Bromatologia i Cheemia Toksykologiczna., 7(161), 1974
23673837tmousetLD50toralt2643 mg/kgt BEHAVIORAL: SOMNOLENCE (GENERAL DEPESSED Activity)t Drug and Chemical Toxicology., 1(89), 1977/1978
23673837tmousetLD50tintravenoust60 mg/kgt Journal of the American Pharmaceutical Association, Scientific Edition., 38(428), 1949

[1]. Preclinical evaluation of docusate as protective agent from herpes simplex viruses. Antiviral Res. 2001 Oct;52(1):25-32.

[2]. Increased adiposity, inflammation, metabolic disruption and dyslipidemia in adult male offspring of DOSS treated C57BL/6 dams. Sci Rep. 2019 Feb 6;9(1):1530.

Sodium docusate is an organic sodium salt. Sodium docusate is the sodium salt of docusate, a dioctyl salt with a skin-softening and laxative effect. Docusate reduces the surface tension and emulsifies the stool, allowing water to penetrate and mix with it, thus softening the stool. It is a versatile surfactant, wetting agent, and solubilizer widely used in the pharmaceutical, cosmetic, and food industries. It has also been used in laxatives and cerumen dissolving agents. It is usually administered in the form of calcium, potassium, or sodium salts. See also: Docusate (contains the active ingredient); Sodium docusate; Sennoside (one of the ingredients); Benzocaine; Sodium docusate (one of the ingredients)... See more...

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Prevention of sexually transmitted infections (STIs) is key to public health control of these diseases. An effective vaginal antimicrobial agent can provide localized, broad-spectrum prevention of the spread of a variety of STI pathogens. Sodium docusate is a sulfated surfactant, and therefore it may inactivate viral pathogens by disrupting the viral envelope and/or denaturing/dissociating proteins. Therefore, this study evaluated the in vitro efficacy and toxicity of sodium docusate (sodium dioctyl sulfosuccinate; Zorex; Meditech Pharmaceuticals, Inc., Scottsdale, Arizona) against herpes simplex virus (HSV). In vitro experiments showed that sodium docusate was effective against both wild-type and resistant strains of HSV types 1 and 2, with an EC(90–100) (effective concentration for 90–100% reduction in viral yield) of approximately 0.005% (w/v). Sodium dodecyl sulfate (SDS) was comparable in potency to sodium docusate, but after 2 days of incubation, sodium docusate showed slightly lower toxicity to uninfected Vero cells compared to SDS (sodium docusate CC(50) approximately 0.01%, while SDS was approximately 0.005%). The cytotoxicity of Sodium docusate is time- and dose-dependent, and therefore related to the frequency of use. The inactivation kinetics were studied by time-course experiments using premixed virus and drug, and the results showed that Sodium docusate could reach its EC (90-100) within 30 minutes. Pretreatment with Sodium docusate reduced the infectivity of cells by 45% despite three washes. Post-infection plaques were reduced by about 30% after treatment. [1]

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Surfactants/detergents are important vaginal antimicrobial agents for controlling sexually transmitted infections. These products destroy microorganisms by dissolving their outer membranes, thereby reducing their infectivity. The study described in this article aimed to examine the antiviral activity of Sodium docusate and its potential role as a topical antimicrobial agent in the prevention of sexually transmitted infections or in the topical treatment of lesions caused by herpes simplex virus. A recent study on nonoxynol-9 as a potential vaginal bactericide (van Damme L., "Advances in Topical Bactericides," presented at the 13th International AIDS Conference in Durban, South Africa, July 9-14, 2000) has raised concerns about the clinical value of surfactants, as nonoxynol-9 users reported a significantly increased transmission rate of 50%, and a direct correlation between frequency of use and HIV transmission risk. Previous analyses of this study did not identify significant clinical toxicity issues with the drug. However, the possibility that cytotoxicity may enhance HIV transmission must be considered. SDS can inactivate various viruses and sexually transmitted infections (Howett et al., 1999; Krebs et al., 1999; Krebs et al., 2000) and is considered a potential topical bactericide. Therefore, we investigated the efficacy of SDS in parallel. We found that docusate at a concentration of 0.005% (w/v) exhibited highly effective killing activity against both HSV-1 and HSV-2 infections, including mutant strains resistant to acyclovir and foscarnet, with an EC90-100 value of approximately 0.005%. While this concentration of SDS was also effective, SDS is cytotoxic, leading to 50% cell death. This result indicates that docusate is less toxic than SDS. As shown in Figure 2, this concentration of docusate reached an EC90-100 value in the HSV-2 inactivation kinetic curve within 30 minutes to 1 hour. Previous studies have found that sodium docusate is effective against a variety of other microorganisms, such as respiratory syncytial virus, influenza virus, bacteria, and fungi (unpublished data from Meditech Pharmaceuticals, Inc.). We hypothesize that it should be effective against all enveloped viruses and possibly also against non-enveloped viruses, as surfactants can denature/dissociate proteins, as demonstrated by sodium dodecyl sulfate (SDS) inactivating non-enveloped human papillomavirus (Howett et al., 1999). We confirm that it remains effective in inactivating HSV-2 and blocking viral adsorption/replication in pretreated cells even after three washes to remove sodium docusate. While the mechanism of cell-drug interaction is unclear, we hypothesize that some sodium docusate may bind/adsorb to the cell surface or be absorbed by the cells. The mild antiviral effect of sodium docusate observed in HSV-infected cells suggests that cellular uptake of sodium docusate may be limited. Surfactants used as topical bactericides generally present cytotoxicity issues due to their lack of selectivity for cell membranes and viral envelopes. This is related to the frequency of use. We have demonstrated that both sodium docusate and sodium dodecyl sulfate (SDS) exhibit time- and dose-dependent effects. We confirm that the viability of cultured cells in vitro depends on the concentration of the bactericide and the duration of exposure. During short-term exposure (1 hour), both sodium docusate and SDS tested showed very low toxicity. During long-term exposure, sodium docusate led to decreased cell viability with increasing concentration. The toxicity of sodium docusate at a concentration of 0.01% in Vero cells reached 50-100% after 3 days of incubation. Sodium docusate was less cytotoxic than SDS. The CC50 of sodium docusate after 2 days of incubation was approximately 0.01%, while that of SDS was approximately 0.005%. The study by Krebs et al. (1999) showed that SDS had the lowest toxicity among N-9, C31G and SDS. This may suggest that sodium docusate is less toxic than N-9 and C31G. Our study suggests that sodium docusate has good in vitro properties and is worth considering as a potent topical antimicrobial agent for the prevention of sexually transmitted infections. The clinical implications of the toxic potential of surfactants in this regard remain to be determined. [1] In conclusion, this work provides evidence of DOSS as a developmental obesity agent in vivo. This study highlights the need to develop effective methods for detecting DOSS in human biosamples and to further investigate the health effects of DOSS exposure in populations. Male offspring showed a wide range of outcomes after treatment with sodium docusate (DOSS), with some animals exhibiting a more pronounced fat accumulation effect and others showing higher/lower fat content, including circulating leptin levels. Natural differences between individuals (e.g., social class) may be a significant factor contributing to the observed differences. Other factors that may contribute to the differences include variations in maternal water intake leading to changes in effective DOSS dose levels, differences in pups’ food intake and physical activity, and the seasonality of offspring birth. Further research is needed to determine the impact of each factor on the differences. However, given the observed metabolic disturbances in male offspring born to DOSS-treated mothers, the safety and prescription of sodium docusate (a stool softener) during pregnancy, as well as potential important dietary sources of DOSS, should be further examined. [2]

C20H37NAO7S

444.5584

444.215

C, 54.04; H, 8.39; Na, 5.17; O, 25.19; S, 7.21

577-11-7

10041-19-7 (acid);128-49-4 (calcium);577-11-7 (sodium);7491-09-0 (potassium); 15968-85-1 (aluminum);

23673837

White to off-white solid powder

1.1

173-179 °C(lit.)

199ºC

4.89

0

7

18

29

546

0

CCCCC(CC)COC(=O)CC(C(=O)OCC(CC)CCCC)S(=O)(=O)[O-].[Na+]

APSBXTVYXVQYAB-UHFFFAOYSA-M

InChI=1S/C20H38O7S.Na/c1-5-9-11-16(7-3)14-26-19(21)13-18(28(23,24)25)20(22)27-15-17(8-4)12-10-6-2;/h16-18H,5-15H2,1-4H3,(H,23,24,25);/q;+1/p-1

sodium;1,4-bis(2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate

Docusate sodium; Dioctylal; Diotilan; Disonate; Molatoc; Regutol; Velmol; ...; 577-11-7; Dioctyl sodium sulfosuccinate (DOSS)

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ≥ 20 mg/mL (~44.99 mM)

Solubility in Formulation 1: 50 mg/mL (112.47 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)